The aim of this project is to determine if an assay using ferromagnetic dense particles (FDP) can improve the detection of the fucosylated glycoforms of secreted liver biomarkers that are present only at very low levels in early-stage liver cancer (HCC) patient serum. Advantages of the FDP format include: very high protein binding capacity, very high mixing efficiency, very rapid reaction kinetics, minimal non-specific interference, and the ability to use unprocessed whole blood samples in a commercial clinical setting. FDP is expected to avoid loss/degradation of rare biomarker species during sample processing and eliminate non-specific interference. The results will be a rapid, accurate, easy to use screening test with a minimal number of processing steps. Thus the aims of this phase I SBIR are to develop a system to neutralize the interference of heterophilic anti-gal antibodies and to develop and optimize an FDP-based assay for three fucosylated biomarkers of HCC using archived serum samples. In Phase II we will further optimize the test for use with unprocessed whole blood and generate a simple to use, rapid and sensitive HCC test to satisfy an important medical need to recognize this highly lethal disease at the earliest possible time. Our commercial objective is to license the validated assay to a clinical diagnostics company that can adapt our FDP-based test to a commercial clinical format, obtain regulatory approval, market and distribute the first reliable and accurate early-detection assay for HCC to the healthcare market. We firmly believe that this phase I proposal will develop an assay robust enough to be used for high throughput screening of patients for the early onset of HCC based on our previous work. Thus, at the conclusion of this project, we strongly believe that we will have developed an FDP-based assay format with our three biomarkers that will allow us to detect liver cancer with >95% sensitivity and >95% specificity, which is substantially better than the current marker AFP, when used alone. Thus, using FDP separation technology in combination with Dr. Mehta's unique biomarkers, specific whole blood diagnostic tests will be developed to meet this underserved medical need. Aim 1. Develop a high throughput ferromagnetic particle (FDP)-based assay to remove heterophilic anti-gal antibodies from serum (6 months).Aim 2. Develop a high throughput ferromagnetic particle (FDP)-based assay to quantitatively capture and detect serum levels of four fucosylated protein biomarkers that are positively correlated with the early stage development of liver cancer and perform a pre-validation study using a broader patient population. PUBLIC HEALTH RELEVANCE: There is a significant unmet medical need for the early detection of liver cancer (HCC) as 5 year survival rates are extremely low. The proposal outlined in this SBIR grant addresses this need. The combination of Russell Biotech's novel FDP separation technology with the biomarkers identified by Dr. Mehta will lead to very rapid, sensitive diagnostic test(s) for the early detection of this devastating cancer.